Journal of
Plant Breeding and Crop Science

  • Abbreviation: J. Plant Breed. Crop Sci.
  • Language: English
  • ISSN: 2006-9758
  • DOI: 10.5897/JPBCS
  • Start Year: 2009
  • Published Articles: 409

Full Length Research Paper

Optimization and development of regeneration and transformation protocol in Indian mustard using lectin gene from chickpea [Cicer arietinum (L.)]

V. V. Singh1, Vandana Verma1*, Aniruddh K. Pareek1, Monika Mathur1, Rajbir Yadav1, Poonam Goyal1, Ajay Kumar Thakur1, Y. P. Singh1, K. R. Koundal2, K. C. Bansal2, A. K. Mishra1, Arvind Kumar1 and Sandeep Kumar1
  1Directorate of Rapeseed-Mustard Research, Bharatpur, Rajasthan 321 303, India. 2NRCPB, IARI, New Delhi, India.  
Email: [email protected], [email protected]

  •  Accepted: 15 October 2009
  •  Published: 30 November 2009

Abstract

 

A simple and easy protocol for regeneration and genetic transformation ofBrassica juncea (L.) Czern and Coss cv. Pusa bold has been developed. The optimum regeneration was found on combination of BAP (3.0 mg/l) and IAA (0.2 mg/l). The genetic transformation protocol has been standardized usingAgrobacterium tumefaciens (gv3101). Cotyledonary petioles used as explant were pre-cultured for 48 h and than co-cultivated with Agrobacterium suspension for 2 days. Putative transformants were selected from selection medium containing 20 mg/l Kanamycine and 250 mg/l cefotaxime. Complete plant formed after placing shoots on rooting medium (0.5 mg/l IBA). Confirmation of integration of transgene was done by PCR using nptII gene primer.

 

Key words: Indian mustard, Agrobacterium tumefaciens, chickpea lectin gene, cotyledonary petioles, transformation.